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Spontaneous, solvent-free entrapment of siRNA within lipid nanoparticles.
Kulkarni, Jayesh A; Thomson, Sarah B; Zaifman, Josh; Leung, Jerry; Wagner, Pamela K; Hill, Austin; Tam, Yuen Yi C; Cullis, Pieter R; Petkau, Terri L; Leavitt, Blair R.
  • Kulkarni JA; NanoMedicines Innovation Network, Vancouver, British Columbia, Canada. pieterc@mail.ubc.ca j.kulkarni@alumni.ubc.ca.
Nanoscale ; 12(47): 23959-23966, 2020 Dec 21.
Article in English | MEDLINE | ID: covidwho-947558
Semantic information from SemMedBD (by NLM)
1. Nucleic Acids COEXISTS_WITH Buffers
Subject
Nucleic Acids
Predicate
COEXISTS_WITH
Object
Buffers
2. P4HTM gene|P4HTM COEXISTS_WITH Nucleic Acids
Subject
P4HTM gene|P4HTM
Predicate
COEXISTS_WITH
Object
Nucleic Acids
3. Nucleic Acids COEXISTS_WITH Buffers
Subject
Nucleic Acids
Predicate
COEXISTS_WITH
Object
Buffers
4. P4HTM gene|P4HTM COEXISTS_WITH Nucleic Acids
Subject
P4HTM gene|P4HTM
Predicate
COEXISTS_WITH
Object
Nucleic Acids
ABSTRACT
Lipid nanoparticle (LNP) formulations of nucleic acid are leading vaccine candidates for COVID-19, and enabled the first approved RNAi therapeutic, Onpattro. LNPs are composed of ionizable cationic lipids, phosphatidylcholine, cholesterol, and polyethylene glycol (PEG)-lipids, and are produced using rapid-mixing techniques. These procedures involve dissolution of the lipid components in an organic phase and the nucleic acid in an acidic aqueous buffer (pH 4). These solutions are then combined using a continuous mixing device such as a T-mixer or microfluidic device. In this mixing step, particle formation and nucleic acid entrapment occur. Previous work from our group has shown that, in the absence of nucleic acid, the particles formed at pH 4 are vesicular in structure, a portion of these particles are converted to electron-dense structures in the presence of nucleic acid, and the proportion of electron-dense structures increases with nucleic acid content. What remained unclear from previous work was the mechanism by which vesicles form electron-dense structures. In this study, we use cryogenic transmission electron microscopy and dynamic light scattering to show that efficient siRNA entrapment occurs in the absence of ethanol (contrary to the established paradigm), and suggest that nucleic acid entrapment occurs through inversion of preformed vesicles. We also leverage this phenomenon to show that specialized mixers are not required for siRNA entrapment, and that preformed particles at pH 4 can be used for in vitro transfection.
Subject(s)

Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Small Interfering / Nanoparticles / Lab-On-A-Chip Devices / SARS-CoV-2 / COVID-19 / Lipids Topics: Vaccines Limits: Animals Language: English Journal: Nanoscale Year: 2020 Document Type: Article

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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Small Interfering / Nanoparticles / Lab-On-A-Chip Devices / SARS-CoV-2 / COVID-19 / Lipids Topics: Vaccines Limits: Animals Language: English Journal: Nanoscale Year: 2020 Document Type: Article